Identification and characterization of de novo-synthesized porcine oviductal secretory proteins.
Oviductal secretory products provide a biochemical environment important for establishment of pregnancy. A previous study identified three de novo-synthesized glycoproteins by one-dimensional SDS-PAGE as well as increased incorporation of [3H]Leu into secretory protein by whole oviduct and ampulla associated with proestrus, estrus, and metestrus only. Here, our objective was to further identify and characterize oviductal secretory proteins, specifically 115,000- and 85,000-Mr estrus-associated proteins (EAP). Two-dimensional SDS-PAGE resolved the 115,000-Mr protein into two proteins of 100,000 Mr, one basic and one acidic, and the 85,000-Mr protein into 75,000- and 85,000-Mr species (pI less than 4.0). Differential secretion of proteins between ampulla and isthmus was indicated. The 100,000-, 75,000-, and 85,000-Mr proteins were synthesized by ampulla during estrus but not by isthmus nor by uterine endometrium. De novo-synthesized EAP were labeled with glucosamine, Leu, and Met, and the 75,000-85,000-Mr proteins from ampulla and a 30,000-Mr family from isthmus were labeled with fucose. Inorganic [35S]sulfate labeled three EAP. Fractionation of culture medium by gel filtration demonstrated differences between products secreted by ampulla and isthmus and suggested that some EAP may be found as high-molecular weight forms in the native state. Results indicate that porcine oviductal tissue synthesizes specific EAP at the time of fertilization and early cleavage-stage embryonic development, that there are differences in the type and distribution of glycoproteins from ampulla and isthmus, and that post-translational modifications occur with the addition of glucosamine, fucose, and inorganic sulfate.
Buhi
W C
WC
Department of Obstetrics and Gynecology, University of Florida, Gainesville 32610.
Alvarez
I M
IM
Sudhipong
V
V
Dones-Smith
M M
MM
D000818:Animals; D003470:Culture Media; D015180:Electrophoresis, Gel, Two-Dimensional; D004717:Endometrium / Q000378:metabolism; D005260:Female; D006023:Glycoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D008970:Molecular Weight; D010057:Oviducts / Q000033:anatomy & histology / Q000378:metabolism*; D014176:Protein Biosynthesis; D011498:Protein Precursors / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D013552:Swine
D003470:Culture Media; D006023:Glycoproteins; D011498:Protein Precursors; D011506:Proteins